The proposed research represents an interdisciplinary effort to understand the pathways for the biosynthesis and metabolism of the fatty acid amide family of bioactive lipids, with a particular focus on a novel set of N-acyltransferases. The N-acyltransferases catalyze the following reaction: acyl-CoA + amine -> N-acylamine + CoA-SH. We have largely completed an evaluation of the endogenous fatty acid amidome in neuroblastoma cells and in Drosophila and need to complete our measurements of fatty acid amidome in choroid plexus cells. Once the baseline fatty acid amide lipidome is understood in these model systems, we will eliminate specific enzymes of fatty acid amide metabolism and measure the corresponding changes fatty acid amides - subtraction lipidomics. More detailed information will come from competitive pulse chase experiments. Over the past 20 years, it has become increasingly clear that the fatty acid amides are a broad family of cell-signaling lipids tremendously important to human health. The members of the fatty acid amide family include the N-acylethanolamines (NAEs), the N-acyldopamines (NADAs), the N-acyltaurines (NATs), N-acylamino acids (NAMAs), and the primary fatty acid amides (PFAMs). Recent studies have suggested that members of the fatty acid amide family are metabolically linked. Our goal is to better define the metabolic relationships between the members of the fatty acid amide family and elaborate the enzymes catalyzing these conversions. A full understanding of fatty acid amide metabolism and the enzymes involved is essential to ultimately developing new drugs and diagnostic strategies to treat human diseases related to fatty acid amide dysfunction.